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Flow cytometry for meningeal infiltration in B acute lymphoblastic leukemia in a low middle income country
Eya Anane1,2, Fatma Ben Lakhal1,2, Sarra Fekih Salem1
1Biological hematology department, Aziza Othmana University Hospital, Tunis, Tunisia.
Insights
Multiparametric flow cytometry (MFC) offers improved detection of meningeal infiltration in pediatric B acute lymphoblastic leukemia compared to conventional cytology. This method enhances diagnostic accuracy, especially in challenging hypocellular cerebrospinal fluid samples, improving patient outcomes.
Area of Science:
- Pediatric Hematology
- Oncology
- Clinical Diagnostics
Background:
- Meningeal infiltration is a severe complication in pediatric B acute lymphoblastic leukemia (B-ALL).
- Early diagnosis is critical for effective treatment, improved outcomes, and reduced relapse rates, particularly in resource-limited settings.
- Conventional cytology (CC) of cerebrospinal fluid (CSF) is the standard but has low sensitivity (<50%).
Purpose of the Study:
- To evaluate the analytical performance of multiparametric flow cytometry (MFC) against conventional cytology (CC) for detecting meningeal infiltration in pediatric B-ALL.
- To assess the utility of MFC as an alternative or complementary diagnostic tool.
Main Methods:
- A cross-sectional study analyzed 60 CSF samples from 39 children with B-ALL over six months.
- CSF samples were analyzed using both CC and MFC for blast cell detection.
- Statistical analysis included Kappa index and chi-squared test to compare methods.
Main Results:
- Meningeal infiltration was detected in 20% of cases by MFC versus 5% by CC (p=0.006).
- The Kappa coefficient was 0.35, indicating weak agreement between CC and MFC.
- MFC demonstrated higher positivity rates, particularly in hypocellular samples (8 positive by MFC vs. 0 by CC).
Conclusions:
- Multiparametric flow cytometry (MFC) exhibits superior sensitivity and good specificity for detecting meningeal involvement in pediatric B-ALL compared to conventional cytology.
- MFC can serve as a valuable complementary method to CC for CNS blast detection, especially in challenging sample types.
Abstract:
Meningeal infiltration in children with B acute lymphoblastic leukemia is one of the most serious complications. Timely diagnosis not only significantly enhances treatment efficacy but also leads to improve patient outcome and reduce risk of relapse. This is particularly crucial in low to middle income countries facing health constraints, where optimizing resources is essential. Conventional cytology (CC) study of cerebrospinal fluid (CSF) is considered in different countries to be the Gold-standard despite its low sensitivity (< 50%). The study of CSF by multiparametric flow cytometry (MFC) appears to be an alternative. The aim of our study was to assess MFC analytical performance compared with CC. Our cross sectional study was conducted over a six-month period in the biological hematology department. CSF samples underwent analysis for the presence of blasts using both CC and MFC. Cytological slides of the CSF were prepared by cytocentrifugation in a Shandon Cytospin 4™. Flow cytometric analysis was performed on the BD FACSLyric™ flow cytometer. All statistical analyses were performed using SPSS version 21.0 (SPSS Inc.). Agreement between the two methods was made using the Kappa index and χ2 test. This study was approved by the local ethics committee. Sixty CSF samples from 39 children with B acute lymphoblastic leukemia were analyzed. Meningeal infiltration was detected respectively in 20% of cases by MFC and 5% of cases by CC, with a significant difference p = 0.006. Comparing the two methods, the Kappa coefficient was 0.35, indicating weak agreement between the two methods. Moreover, MFC positivity was higher even for hypocellular samples. Of the 51 hypocellular samples, eight were positive by MFC while they were negative by CC. MFC shows better sensitivity while retaining good specificity for the detection of meningeal involvement. MFC could therefore be a complementary method to CC for detecting blast cells in the central nervous system.
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